#METABOLOMICS WORKBENCH juliehaines_20241126_073648 DATATRACK_ID:5405 STUDY_ID:ST003610 ANALYSIS_ID:AN005934 PROJECT_ID:PR002231 VERSION 1 CREATED_ON December 2, 2024, 3:15 pm #PROJECT PR:PROJECT_TITLE Macrophages recycle phagocytosed bacteria to fuel immunometabolic responses PR:PROJECT_SUMMARY Macrophages specialize in phagocytosis, a cellular process that eliminates PR:PROJECT_SUMMARY extracellular matter, including microbes, through internalization and PR:PROJECT_SUMMARY degradation. Despite the critical role of phagocytosis during bacterial PR:PROJECT_SUMMARY infection, the fate of phagocytosed microbial cargo and its impact on host cell PR:PROJECT_SUMMARY is poorly understood. Here, we reveal that ingested bacteria constitute an PR:PROJECT_SUMMARY alternative nutrient source that skews immunometabolic host responses. Tracing PR:PROJECT_SUMMARY stable isotope-labelled bacteria, we found that phagolysosomal degradation of PR:PROJECT_SUMMARY bacteria provides carbon atoms and amino acids that are recycled into various PR:PROJECT_SUMMARY metabolic pathways, including glutathione and itaconate biosynthesis, and PR:PROJECT_SUMMARY satisfy macrophage bioenergetic needs. Metabolic recycling of PR:PROJECT_SUMMARY microbially-derived nutrients is regulated by the nutrient sensing mTORC1 and PR:PROJECT_SUMMARY intricately tied to microbial viability. Dead bacteria, as opposed to live ones, PR:PROJECT_SUMMARY are enriched in cyclic- adenosine monophosphate (AMP), sustain the cellular AMP PR:PROJECT_SUMMARY pool and subsequently activate AMP protein kinase (AMPK) to inhibit mTORC1. PR:PROJECT_SUMMARY Consequently, killed bacteria strongly fuel metabolic recycling and support PR:PROJECT_SUMMARY macrophage survival, but elicit decreased reactive oxygen species (ROS) PR:PROJECT_SUMMARY production and a reduced IL-1β secretion compared to viable bacteria. These PR:PROJECT_SUMMARY results reveal a novel insight into the fate of engulfed microbes and highlights PR:PROJECT_SUMMARY a microbial viability-associated metabolite that triggers host metabolic and PR:PROJECT_SUMMARY immune responses. Our findings hold promise for shaping immunometabolic PR:PROJECT_SUMMARY intervention in various immune-related pathologies. PR:INSTITUTE University of Colorado Anschutz Medical Campus PR:LABORATORY Lab of Angelo D'Alessandro in collaboration with lab of Johan Garaude (INSERM, PR:LABORATORY Fr) PR:LAST_NAME Haines PR:FIRST_NAME Julie PR:ADDRESS 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA PR:EMAIL julie.haines@cuanschutz.edu PR:PHONE 3037243339 #STUDY ST:STUDY_TITLE 13C-tracing metabolomics of peritoneal macrophages 1hour and 2hours after ST:STUDY_TITLE engulfment of 13C-labeled heat-killed E coli ST:STUDY_SUMMARY 13C-tracing analysis at 1h and 2h of peritoneal macrophages isolated from ST:STUDY_SUMMARY C57BL6/N mice injected with uniformly 13C-labelled and DAPI labeled heat-killed ST:STUDY_SUMMARY E. coli. ST:INSTITUTE University of Colorado Anschutz Medical Campus ST:LAST_NAME Haines ST:FIRST_NAME Julie ST:ADDRESS 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA ST:EMAIL julie.haines@cuanschutz.edu ST:PHONE 3037243339 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Mus musculus SU:GENDER Not applicable #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - AA83PM-1_r19- Killed E coli labeling:13C-labelled KEC | Timepoint:1h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-1_r19- SUBJECT_SAMPLE_FACTORS - AA83PM-2_r10- Killed E coli labeling:13C-labelled KEC | Timepoint:1h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-2_r10- SUBJECT_SAMPLE_FACTORS - AA83PM-3_r17- Killed E coli labeling:13C-labelled KEC | Timepoint:1h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-3_r17- SUBJECT_SAMPLE_FACTORS - AA83PM-4_r15- Killed E coli labeling:13C-labelled KEC | Timepoint:1h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-4_r15- SUBJECT_SAMPLE_FACTORS - AA83PM-5_r9- Killed E coli labeling:13C-labelled KEC | Timepoint:1h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-5_r9- SUBJECT_SAMPLE_FACTORS - AA83PM-6_r3- Killed E coli labeling:Unlabeled KEC | Timepoint:1h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-6_r3- SUBJECT_SAMPLE_FACTORS - AA83PM-7_r16- Killed E coli labeling:Unlabeled KEC | Timepoint:1h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-7_r16- SUBJECT_SAMPLE_FACTORS - AA83PM-8_r5- Killed E coli labeling:Unlabeled KEC | Timepoint:1h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-8_r5- SUBJECT_SAMPLE_FACTORS - AA83PM-9_r13- Killed E coli labeling:Unlabeled KEC | Timepoint:1h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-9_r13- SUBJECT_SAMPLE_FACTORS - AA83PM-10_r6- Killed E coli labeling:13C-labelled KEC | Timepoint:2h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-10_r6- SUBJECT_SAMPLE_FACTORS - AA83PM-11_r2- Killed E coli labeling:13C-labelled KEC | Timepoint:2h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-11_r2- SUBJECT_SAMPLE_FACTORS - AA83PM-12_r18- Killed E coli labeling:13C-labelled KEC | Timepoint:2h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-12_r18- SUBJECT_SAMPLE_FACTORS - AA83PM-13_r7- Killed E coli labeling:13C-labelled KEC | Timepoint:2h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-13_r7- SUBJECT_SAMPLE_FACTORS - AA83PM-14_r8- Killed E coli labeling:13C-labelled KEC | Timepoint:2h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-14_r8- SUBJECT_SAMPLE_FACTORS - AA83PM-15_r12- Killed E coli labeling:Unlabeled KEC | Timepoint:2h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-15_r12- SUBJECT_SAMPLE_FACTORS - AA83PM-16_r14- Killed E coli labeling:Unlabeled KEC | Timepoint:2h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-16_r14- SUBJECT_SAMPLE_FACTORS - AA83PM-17_r4- Killed E coli labeling:Unlabeled KEC | Timepoint:2h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-17_r4- SUBJECT_SAMPLE_FACTORS - AA83PM-18_r11- Killed E coli labeling:Unlabeled KEC | Timepoint:2h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-18_r11- SUBJECT_SAMPLE_FACTORS - AA83PM-19_r1- Killed E coli labeling:Unlabeled KEC | Timepoint:2h | Sample source:cells RAW_FILE_NAME(raw file name)=AA83PM-19_r1- #COLLECTION CO:COLLECTION_SUMMARY Cells from the peritoneal cavity were harvested with cold PBS. Phagocytic cells CO:COLLECTION_SUMMARY (DAPI+ cells) were sorted by flow cytometry, frozen as dry cell pellet, and CO:COLLECTION_SUMMARY stored at -80˚C until processing. CO:SAMPLE_TYPE Macrophages #TREATMENT TR:TREATMENT_SUMMARY Preparation of macrophages: Mice were injected or not with 0.5 ml of TR:TREATMENT_SUMMARY thioglycolate medium to increase peritoneal macrophage abundance. Four days TR:TREATMENT_SUMMARY later, mice were injected with 1E9 killed U-[13C]Bacteria previously labeled TR:TREATMENT_SUMMARY with DAPI. One or two hours later, peritoneal cells were harvested, and sorted TR:TREATMENT_SUMMARY by flow cytometry based on DAPI. Preparation of viable and killed TR:TREATMENT_SUMMARY U-[13C]Bacteria: ThyA- E. coli were grown overnight with shaking in LB TR:TREATMENT_SUMMARY supplemented with thymidine (500 ug/ml) and trimethoprim (50 ug/ml), diluted TR:TREATMENT_SUMMARY 1/40, and grown until log-phase [optical density at 600 nm (OD600) of 0.8-1.2]. TR:TREATMENT_SUMMARY Bacteria were washed with phosphate buffer saline (PBS) to remove LB salts TR:TREATMENT_SUMMARY before addition to cells. For labeling of bacteria, 10 ul of an overnight TR:TREATMENT_SUMMARY cultured of thyA- E. coli was added to 20 ml of a filtered M9 minimal medium TR:TREATMENT_SUMMARY salts (Life Technologies) supplemented with 1 mM thiamine, 1 mM MgSO4, 0.1 M TR:TREATMENT_SUMMARY CaCl2, 500 ug/ml thymidine, 50 ug/ml trimethoprim, and 0.5% U-[13C] glucose TR:TREATMENT_SUMMARY (Campro Scientific). Bacteria were grown for 72h, washed with PBS and subjected TR:TREATMENT_SUMMARY to heat-killing by re-suspension in PBS and subsequently incubation at 60˚C for TR:TREATMENT_SUMMARY 60-90 min. Bacteria were kept at 4˚C until use. Efficient killing was confirmed TR:TREATMENT_SUMMARY by overnight plating on LB-agar plates. For DAPI labelling, killed bacteria were TR:TREATMENT_SUMMARY incubated with 0.2 ug/ml of DAPI in PBS for 5min and washed 3 times with cold TR:TREATMENT_SUMMARY PBS. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Metabolites from frozen pellets were extracted at 4e6 cells per mL using ice SP:SAMPLEPREP_SUMMARY cold 5:3:2 methanol:acetonitrile:water (v/v/v) with vigorous vortexing at 4 SP:SAMPLEPREP_SUMMARY degrees C followed by centrifugation as described for 10 min at 18,000 g. SP:SAMPLEPREP_SUMMARY Supernatants were maintained at 4°C until analysis that same day. SP:PROCESSING_STORAGE_CONDITIONS 4℃ SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Positive C18 CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Phenomenex Kinetex C18 (150 x 2.1mm,1.7um) CH:SOLVENT_A 100% water; 0.1% formic acid CH:SOLVENT_B 100% acetonitrile; 0.1% formic acid CH:FLOW_GRADIENT 0-0.5 min 5% B, 0.5-1.1 min 5-95% B, 1.1-2.75 min hold at 95% B, 2.75-3 min CH:FLOW_GRADIENT 95-5% B, 3-5 min hold at 5% B CH:FLOW_RATE 450 uL/min CH:COLUMN_TEMPERATURE 45 CH:SAMPLE_INJECTION 10 uL #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Orbitrap Exploris 120 MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS We use a Thermo Orbitrap Exploris 120. Resolution 120,000, scan range 65-975 MS:MS_COMMENTS m/z, maximum injection time 100 ms, microscans 1, automatic gain control (AGC) MS:MS_COMMENTS detection duration 20 msec, source voltage 3.5 kV, capillary temperature 320 C, MS:MS_COMMENTS vaporizer temp 200 C, and sheath gas 50, auxiliary gas 10, and sweep gas 1 (all MS:MS_COMMENTS nitrogen). Data converted to mzXML using RawConverter. Metabolites were MS:MS_COMMENTS annotated and integrated using Maven in conjunction with the KEGG database. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples AA83PM-1_r19- AA83PM-2_r10- AA83PM-3_r17- AA83PM-4_r15- AA83PM-5_r9- AA83PM-6_r3- AA83PM-7_r16- AA83PM-8_r5- AA83PM-9_r13- AA83PM-10_r6- AA83PM-11_r2- AA83PM-12_r18- AA83PM-13_r7- AA83PM-14_r8- AA83PM-15_r12- AA83PM-16_r14- AA83PM-17_r4- AA83PM-18_r11- AA83PM-19_r1- Factors Killed E coli labeling:13C-labelled KEC | Timepoint:1h | Sample source:cells Killed E coli labeling:13C-labelled KEC | Timepoint:1h | Sample source:cells Killed E coli labeling:13C-labelled KEC | Timepoint:1h | Sample source:cells Killed E coli labeling:13C-labelled KEC | Timepoint:1h | Sample source:cells Killed E coli labeling:13C-labelled KEC | Timepoint:1h | Sample source:cells Killed E coli labeling:Unlabeled KEC | Timepoint:1h | Sample source:cells Killed E coli labeling:Unlabeled KEC | Timepoint:1h | Sample source:cells Killed E coli labeling:Unlabeled KEC | Timepoint:1h | Sample source:cells Killed E coli labeling:Unlabeled KEC | Timepoint:1h | Sample source:cells Killed E coli labeling:13C-labelled KEC | Timepoint:2h | Sample source:cells Killed E coli labeling:13C-labelled KEC | Timepoint:2h | Sample source:cells Killed E coli labeling:13C-labelled KEC | Timepoint:2h | Sample source:cells Killed E coli labeling:13C-labelled KEC | Timepoint:2h | Sample source:cells Killed E coli labeling:13C-labelled KEC | Timepoint:2h | Sample source:cells Killed E coli labeling:Unlabeled KEC | Timepoint:2h | Sample source:cells Killed E coli labeling:Unlabeled KEC | Timepoint:2h | Sample source:cells Killed E coli labeling:Unlabeled KEC | Timepoint:2h | Sample source:cells Killed E coli labeling:Unlabeled KEC | Timepoint:2h | Sample source:cells Killed E coli labeling:Unlabeled KEC | Timepoint:2h | Sample source:cells Arginine 2004916.38 3708315.25 2534091.25 1793103.5 3043012 1097618.25 700492.81 744199 1201608.88 4224471.5 5195096.5 4143589.75 3546892.75 3575247 3165262.75 1843420 1198579.38 2610918 1526788.38 Arginine 13C1 680.57892 0 0 0 0 1179.2855 0 0 0 0 0 0 0 3186.588 0 0 0 0 0 Arginine 13C2 0 2385.378771 0 0 0 941.214903 8653.80332 575.399156 0 0 2209.219646 6834.583049 0 0 0 0 0 0 0 Arginine 13C3 0 0 0 0 0 2884.27 0 0 0 0 6752.87 4518.86 4093.39 0 1695.63 0 0 0 0 Arginine 13C4 0 6462.05 0 0 0 0 0 0 0 14997.85 23624.06 34671.08 14641.84 7266.78 0 0 0 1902.08 0 Arginine 13C5 167888.33 248388.98 151414.27 69192.99 182786.64 0 0 5125.04 7424.59 51221.37 86599.84 80170.98 43358.99 57181.5 44095.94 3406.42 7488.79 3151.25 2841.69 Arginine 13C6 37971.5 25656.83 0 10176.61 59851.06 0 0 4338.32 0 136738.58 262318.53 248414.58 165070.75 147381.12 8788.3 9461.12 19865.08 8933.17 14457.02 Cysteine 958682 1875731.5 1956012.5 951312.56 1646443.62 0 0 4530.57 15411.97 1480398.12 1381871 1361953.25 956609.81 800910.31 1320729.88 458879.12 522310.03 315993.09 390799 Cysteine 13C1 19447.28 45369.68 50307.74 24647.67 42950.2 0 0 0 0 41498.45 38097.55 47502.64 27464.86 21332.67 36829.49 13764.18 13581.63 5017.07 13866.53 Glutamate 2772581.75 4890240.5 7132954.5 3199696.75 4547700 340326.62 256446.92 280798.59 513448.53 3158277.25 2883424 2743184.75 2183009 2287379.25 5783524 3207307.75 3265406.25 2301594.25 3212858.25 Glutamate 13C1 201405.6638 190383.8925 0 0 132839.22 0 0 0 0 62432.00125 206191.43 185826.9988 167987.095 116314.0913 39842.3 51706.18375 66344.60625 28544.75625 79794.71625 Glutamate 13C2 87948.16021 95792.90249 21607.59264 11701.96733 77237.0575 0 0 1980.184265 2240.638197 81757.66132 238273.7824 187679.3661 200801.6678 125303.5678 31960.3799 43326.85406 78240.04609 39606.98739 66499.65379 Glutamate 13C3 117297.79 112675.75 4712.88 7271.54 125893.46 0 1952.79 2701.6 0 150765.42 544977.81 339686.72 421367.5 334994.47 5988.69 23466.29 57331.33 20424.62 32084.62 Glutamate 13C4 159371.27 316309.72 260699.17 94916.77 186769.88 0 0 0 0 112164.37 142964.33 110832.94 85937.07 96370.62 74985.69 14916.73 18198.84 11504.82 15774.17 Glutamate 13C5 45396.44 75478.08 0 3536.11 53605.15 0 0 0 0 212245.3 611699.56 392990.91 626669.25 521432.41 14243.95 26586.91 54338.75 17140.06 31332.73 Glutamine 858500 1568696.62 1856303.12 1018429 1431801.62 31542.08 35240.76 35572.19 80065.46 3184359.25 3259919 3429034 2604102.75 2709811 2677590.75 1307775.38 1421825.38 1682367.38 1460185 Glutamine 13C1 0 0 0 0 0 0 517.6682 0 0 0 0 7304.15 4456.81875 0 0 0 0 0 0 Glutamine 13C2 1229.0175 0 0 1522.582275 0 1928.325208 0 1911.664125 871.3719835 10951.12327 15013.90503 19635.36215 12625.98918 10631.90173 4069.087981 1576.479476 4840.908226 0 0 Glutamine 13C3 4464.84 18790.69 0 0 0 0 0 0 0 25208.5 42574.04 50144.08 54814.89 40618.66 0 0 0 0 1980.8 Glutamine 13C4 0 0 0 0 3482.46 0 0 1567.65 0 16843.97 30318.57 25470.06 22686.34 25087.63 0 0 0 0 0 Glutamine 13C5 43097.84 107344.16 63133.19 56287.64 75425.34 0 0 0 7164.18 147635.02 346046.22 356439.38 372703.03 315693.81 33338.92 17973.76 40137.45 3971.72 18558.37 Methionine 4229174.5 8683290 8887882 3201892 5927589.5 81978.26 54730.19 178533.98 317381.22 2488770.5 2480157.25 2119194.25 1602805 1499039.38 2694905.25 831078.5 786391.94 647710.44 817662.5 Methionine 13C1 0 0 0 0 30874.1075 0 41.35955 736.0311 0 0 0 0 4700.425 0 0 0 0 2267.8158 0 Methionine 13C4 16363.47 5862.84 0 0 8712.44 0 1556.4 0 0 17116.43 23699.71 25748.49 28993.34 14983.79 0 0 4863.26 4105.39 0 Methionine 13C5 63225.25 77924.09 0 11455.56 76353.3 0 0 0 0 117078.9 152168.12 197744.5 173935.83 107034.38 5253.5 13107 31756.65 6904.24 15547.11 Adenosine 384789.16 450800.47 413125.97 417695.59 332651.41 147037.38 83951.73 122875.43 84601.55 134478.95 81120.77 165695.19 135486.3 139808.47 175355.31 144400.19 84765.12 152104.42 157829.56 Adenosine 13C1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 7681.1291 0 0 0 Cystathionine 43726 87540.4 36233.2 42005.97 41710.35 0 0 0 11149.14 22375.83 14849.12 17332.05 8027.55 13275.08 38323.66 13578.62 25729.9 23451.85 22591.77 S-Adenosyl-L-methionine 15795.9 68691.83 78382.75 30399.08 37032.07 5456.17 21557.44 5645.03 5120.07 34408.07 33403.74 41397.37 21412.33 39056.22 40635.02 19912.72 29558.99 19116.82 19818.02 Ornithine 1556729.62 3554205.25 3833636.25 1540961.88 1611492.88 61845.7 48191.74 54650.88 53939.07 2545209.75 2094394.88 2591712.25 1876792.12 1519936.62 995583.81 324655.72 631071.44 411298.41 435616.72 Ornithine 13C1 0 0 0 0 0 303.5465 0 0 0 0 0 0 0 0 0 0 0 0 435.6104 Ornithine 13C2 0 0 0 1852.410313 0 0 0 0 0 0 3388.395488 15037.24044 9300.563837 8579.391725 0 1796.676447 0 1565.44231 296.739422 Ornithine 13C3 36493.72 25856.87 0 6800.73 22544.62 0 0 0 4712.07 84346.15 209980.08 216120.5 207603.92 117598.3 2490.46 3435.46 14114.13 3214.56 2544.85 Ornithine 13C4 7304.5 8043.04 0 0 0 0 0 0 0 24740.8 50625.59 54286.79 54459.14 36560.46 0 0 2574.38 0 2318.71 Ornithine 13C5 93080.51 112897.27 0 6605.22 84980.79 0 0 0 0 337708.81 631332.69 621362 699333.44 453153.16 4316.9 8348.3 38697.95 11463.02 24897.83 Putrescine 141001.86 152300.83 48475.5 25057.94 57351.4 12124.06 4631.42 2809.27 8160.6 33910.29 13268.94 11030.57 4271.87 9462.22 14200.11 9165.12 0 30792.63 2331.81 Spermidine 271402.59 338069.84 319612.25 181128.83 178589.69 127953.27 90843.94 71750.98 72583.62 79480.34 24096.62 35010.28 21841.37 15104.24 39049.14 20142.76 34735.98 35647.61 9666.98 Spermine 35177.02 68118.05 68926.96 33562.51 55149.96 25717.3 16326.04 16473.59 7737.56 14758.73 3343.82 3913.14 4519.96 2234.38 11106.49 3044.7 55273.93 5432.84 2455.22 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name KEGG ID mz rt Arginine C00062 175.119049 0.585 Arginine 13C1 C00062 176.122208 0.585 Arginine 13C2 C00062 177.125107 0.604 Arginine 13C3 C00062 178.129425 0.596 Arginine 13C4 C00062 179.131958 0.59 Arginine 13C5 C00062 180.136322 0.648 Arginine 13C6 C00062 181.139053 0.59 Cysteine C00097 122.027214 0.627 Cysteine 13C1 C00097 123.030487 0.627 Glutamate C00025 148.060516 0.612 Glutamate 13C1 C00025 149.063889 0.614 Glutamate 13C2 C00025 150.066528 0.62 Glutamate 13C3 C00025 151.071213 0.611 Glutamate 13C4 C00025 152.074844 0.65 Glutamate 13C5 C00025 153.077194 0.618 Glutamine C00064 147.076492 0.606 Glutamine 13C1 C00064 148.079681 0.611 Glutamine 13C2 C00064 149.081467 0.615 Glutamine 13C3 C00064 150.087128 0.605 Glutamine 13C4 C00064 151.089539 0.641 Glutamine 13C5 C00064 152.092697 0.579 Methionine C00073 150.058441 0.687 Methionine 13C1 C00073 151.061615 0.702 Methionine 13C4 C00073 154.071518 0.712 Methionine 13C5 C00073 155.075394 0.701 Adenosine C00121 268.104218 0.674 Adenosine 13C1 C00121 269.107666 0.683 Cystathionine C00542 223.074661 0.647 S-Adenosyl-L-methionine C00019 399.144531 0.599 Ornithine C00077 133.097214 0.571 Ornithine 13C1 C00077 134.100662 0.572 Ornithine 13C2 C00077 135.102081 0.618 Ornithine 13C3 C00077 136.106491 0.571 Ornithine 13C4 C00077 137.11055 0.568 Ornithine 13C5 C00077 138.113785 0.577 Putrescine C00134 89.107475 0.56 Spermidine C00315 146.165146 0.554 Spermine C00750 203.222992 0.56 METABOLITES_END #END